General Obstetrics and Gynecology: Gynecology
Culture-independent analysis of vaginal microflora: The unrecognized association of Atopobium vaginae with bacterial vaginosis

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Abstract

Although the pathogenesis of bacterial vaginosis remains elusive, a few microorganisms, such as Gardnerella vaginalis, are considered markers on Gram stain or culture. Culture-independent analysis of vaginal microflora using 16S rDNA cloning and sequencing of total bacterial communities reveals the gram-positive Atopobium vaginae as a predominant species in disturbed vaginal flora.

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Material and methods

Detailed information of microbial communities can now be acquired from the phylogenetic analysis of 16S rDNA sequences obtained from clinical samples by polymerase chain reaction (PCR) amplification.2 16S rDNA gene fragments were amplified from a vaginal swab by PCR with universal 16S rDNA primers. The PCR mixture obtained represents all species constituting the vaginal microflora. Within the mixture, each 16S rDNA fragment is unique to a bacterial species, and yields a genomic fingerprint on

Results

Eight healthy nonpregnant women (mean age 41.4 years, range 28 to 51 years) attending our outpatient clinic were selected based on a vaginal Gram stain smear according to Hay et al.4 We included 3 women with healthy (grade I) flora, and 5 women with intermediate (grade II) flora, or with overt bacterial vaginosis (grade III). 16S rDNA was cloned by inserting each 16S rDNA fragment in an Escherichia coli plasmid vector, yielding 100 clones on average (range 70 to 169 clones per patient).

Comment

By cloning and sequencing 16S rDNA we identified a series of species that have not been described before as (common) members of the vaginal community, including Atopobium rimae, Bifidobacterium biavatii (urinalis), Dialister sp., Leptotrichia amnionii, and Sneathia sanguinegens.

In this sample of women of reproductive age, culture-independent analysis also points at a strong correlation between A vaginae and perturbation of the vaginal niche. Phenotypic characterization of this species of

Acknowledgements

This work was part of a research project on determinants of differential susceptibility to ascending genital tract infection, and was supported through a research grant by the Marguerite-Marie Delacroix Foundation. As the main funding source, the Marguerite-Marie Delacroix Foundation was not involved in the development of the study design, the collection, analysis, and interpretation of the data, in the writing of the report, or in the decision to submit the paper for publication. H. V. and M.

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